The integrin-binding defective FGF2 mutants potently suppress FGF2 signalling and angiogenesis

被引:42
作者
Mori, Seiji [1 ,2 ]
Hatori, Nobuaki [2 ]
Kawaguchi, Naomasa [2 ]
Hamada, Yoshinosuke [2 ]
Shih, Tsung-Chieh [3 ]
Wu, Chun-Yi [3 ]
Lam, Kit S. [3 ]
Matsuura, Nariaki [2 ]
Yamamoto, Hirofumi [2 ]
Takada, Yoko K. [3 ,4 ,5 ]
Takada, Yoshikazu [3 ,4 ,5 ]
机构
[1] Morinomiya Univ Med Sci, Fac Hlth Sci, Dept Med Technol, 1-26-16 Nanko Kita, Osaka 5598611, Japan
[2] Osaka Univ, Grad Sch Med, Div Hlth Sci, Dept Mol Pathol, 1-7 Yamadaoka, Suita, Osaka 5650871, Japan
[3] Univ Calif Davis, Sch Med, Dept Biochem & Mol Med, Sacramento, CA 95817 USA
[4] Univ Calif Davis, Sch Med, Dept Dermatol, Sacramento, CA 95817 USA
[5] Taipei Med Univ, Coll Med Sci & Technol, PhD Program Translat Med, 250 Wu Hsing St, Taipei 11031, Taiwan
基金
日本学术振兴会;
关键词
GROWTH-FACTOR RECEPTOR; ALPHA(V)BETA(3); FAMILY; CELLS; ALPHA-V-BETA-3; PROTEIN; REQUIREMENT; ANTAGONIST;
D O I
10.1042/BSR20170173
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We recently found that integrin alpha v beta 3 binds to fibroblast growth factor (FGF)-alpha v beta 31 (FGF1), and that the integrin-binding defective FGF1 mutant (Arg-50 to glutamic acid, R50E) is defective in signalling and antagonistic to FGF1 signalling. R50E suppressed angiogenesis and tumour growth, suggesting that R50E has potential as a therapeutic. However, FGF1 is unstable, and we had to express R50E in cancer cells for xenograft study, since injected R50E may rapidly disappear from circulation. We studied if we can develop antagonist of more stable FGF2. FGF2 is widely involved in important biological processes such as stem cell proliferation and angiogenesis. Previous studies found that FGF2 bound to alpha v beta 3 and antagonists to alpha v beta 3 suppressed FGF2-induced angiogenesis. However, it is unclear how FGF2 interacts with integrins. Here, we describe that substituting Lys-119/Arg-120 and Lys-125 residues in the predicted integrin-binding interface of FGF2 to glutamic acid (the K119E/R120E and K125E mutations) effectively reduced integrin binding to FGF2. These FGF2 mutants were defective in signalling functions (ERK1/2 activation and DNA synthesis) in NIH3T3 cells. Notably they suppressed, FGF2 signalling induced by WT FGF2 in endothelial cells, suggesting that the FGF2 mutants are antagonists. The FGF2 mutants effectively suppressed tube formation in vitro, sprouting in aorta ring assays ex vivo and angiogenesis in vivo. The positions of amino acids critical for integrin binding are different between FGF1 and FGF2, suggesting that they do not interact with integrins in the same manner. The newly developed FGF2 mutants have potential as anti-angiogenic agents and useful tools for studying the role of integrins in FGF2 signalling.
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页数:10
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